EP3160647B1 - Mikrofluidische testkartusche ohne aktive fluidregelung - Google Patents

Mikrofluidische testkartusche ohne aktive fluidregelung Download PDF

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Publication number
EP3160647B1
EP3160647B1 EP15816063.0A EP15816063A EP3160647B1 EP 3160647 B1 EP3160647 B1 EP 3160647B1 EP 15816063 A EP15816063 A EP 15816063A EP 3160647 B1 EP3160647 B1 EP 3160647B1
Authority
EP
European Patent Office
Prior art keywords
test device
sample
reagent
flow paths
well
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
EP15816063.0A
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English (en)
French (fr)
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EP3160647A4 (de
EP3160647A1 (de
Inventor
Manish Deshpande
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Siemens Healthcare Diagnostics Inc
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Siemens Healthcare Diagnostics Inc
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Publication date
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Publication of EP3160647A1 publication Critical patent/EP3160647A1/de
Publication of EP3160647A4 publication Critical patent/EP3160647A4/de
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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/148—Specific details about calibrations
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/087—Multiple sequential chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum

Definitions

  • the invention relates to microfluidic test cartridges for medical diagnostics, and more specifically to tests requiring no onboard fluidic controls and with on-board calibrators.
  • a fluid system in general, may comprise a fluidic device that operates by the interaction of streams of fluid.
  • miniature fluidic devices such as microfluidic devices and biochips
  • a fluidic device in this field usually provides integration of multiple analytical steps into a single device.
  • a fluidic device may perform one or more assays.
  • an assay may be defined as a procedure for quantifying the amount or the functional activity of an analyte in a liquid sample.
  • a typical on-chip assay may involve a variety of on-board operations, such as sample introduction and preparation, metering, sample/reagent mixing, liquid transport, and detection, etc.
  • Typical diagnostic assays involve manipulating very small volumes of fluid with highly precise control.
  • a traditional microfluidic flow device with microfluidic channels, valves and other flow control mechanisms pose specific challenges to ensure the required precision due to several effects including fluid loss in transport, capillary effects, impact of gravity, trapped air and others. Additionally, several assay processes such as mixing and incubation can also pose unique challenges in the microfluidic environment.
  • the ideal choice would be to limit or eliminate the need for flow and flow control, and yet provide the level of precision needed to deliver the required assay performance. This has been accomplished in macro-scale instrumentation, but typically not in a single use disposable format compatible with a small form factor instrument.
  • microfluidic devices require flow to move sample and/or reagents through the disposable from the loading to the detection site. These may use on-board or off-board pumping, capillary or lateral flow, and a variety of fluid control mechanisms, including external valving, mixing methods etc. Precision is typically achieved using appropriate actuation mechanisms. Other sources of potential errors are typically controlled using on- or off-chip components such as bubble traps and capillary barriers.
  • WO-A-2011/071772 discloses assay cartridges and methods of using the same.
  • the invention relates to a test device and an analytical system comprising said test device as defined in the appended claims.
  • This invention includes a device with no active fluid control required on-board. Precision control mechanisms are moved off the disposable to the instrument which allows them to be reusable and therefore potentially more expensive. The consumable is extremely simple and potentially very cost effective as a high-volume disposable.
  • the consumable is a test device for conducting an in-vitro diagnostics test that can be read optically.
  • This may include immunoassays, chemistries, or hematological assays or any other assessment of bodily fluid components that can be analyzed through optical detection.
  • assays that may be carried out during the use of the invention described herein include, but are not limited to, tests for blood gases, clotting factors, immunogens, bacteria, and proteins.
  • the assays that may be detected with the test device is a "luminescent O2 channel assay" (LOCI®) which includes the use of for example, Sandwich Assays based on an analyte-specific antibody and a biotinylated antibody wherein specific wavelengths are generated by the fluid subsample and detected by the test device.
  • Reagent configurations for the assay method include for example Sandwich Formats based on an antigen or an antibody, a Competitive Format, or a Sandwich Format with Extended Linker and may be used in immunoassays, infectious disease testing, and DNA testing.
  • Specific blood chemicals which may be measured include, but are not limited to, TSH, free T4, free T3, Total PSA, free PSA, AFP, CEA, CA15.3, CA 19-9, CA 125, Cardiac Troponin-I, NT-pro BNP, myoglobin, mass CKMB (MMB), BNP, Ferritin, Vitamin B12, Folate, total B-HCG, FSH, LH, prolactin, estradiol, testosterone, progesterone, and digoxin.
  • Fluorescent detection also can be useful for detecting analytes in the presently claimed and disclosed inventive concepts.
  • Useful fluorochromes include, but are not limited to, DAPI, fluorescein, lanthanide metals, Hoechst 33258, R-phycocyanin, B-phycoerythrin, R-phycoerythrin, rhodamine, Texas red and lissamine. Fluorescent compounds, can be chemically coupled to antibodies without altering their binding capacity. When activated by illumination with light of a particular wavelength, the fluorochrome-labelled antibody adsorbs the light energy, inducing a state of excitability in the molecule, followed by emission of the light at a characteristic color visually detectable with a light microscope. Radioimmunoassays (RIAs) can be useful in certain methods of the invention. Such assays are well known in the art. Radioimmunoassays can be performed, for example, with 1251-labeled primary or secondary antibody.
  • Separation steps are possible in which an analyte is reacted with reagent in a first reaction chamber and then the reacted reagent or sample is directed to a second reaction chamber for further reaction.
  • a reagent can be re-suspended in a first reaction chamber and moved to a second reaction chamber for a reaction.
  • An analyte or reagent can be trapped in a first or second chamber and a determination made of free versus bound reagent.
  • the determination of a free versus bound reagent is particularly useful for multizone immunoassay and nucleic acid assays. There are various types of multizone immunoassays that could be adapted to this device.
  • Immunoassays or DNA assay can be developed for detection of bacteria such as Gram negative species (e.g., E. coli, Enterobacter, Pseudomonas, Klebsiella) and Gram positive species (e.g., Staphylococcus aureus, Enterococcus).
  • Immunoassays can be developed for complete panels of proteins and peptides such as albumin, hemoglobin, myoglobulin, ⁇ -1-microglobulin, immunoglobulins, enzymes, glycoproteins, protease inhibitors, drugs and cytokines.
  • the device may be used in analysis of urine for one or more components therein or aspects thereof, such as, but not limited to, leukocytes, nitrites, urobilinogen, proteins, albumin, creatinine, uristatin, calcium oxalate, myoglobin, pH, blood, specific gravity, ketone, bilirubin and glucose.
  • the consumable in non-limiting embodiments, may be made of plastics such as polycarbonate, polystyrene, polyacrylates, or polyurethane, alternatively or in addition to, they can be made from silicates, and/or glass.
  • the plastics preferably used may include, but are not limited to, ABS, acetals, acrylics, acrylonitrile, cellulose acetate, ethyl cellulose, alkylvinylalcohols, polyaryletherketones, polyetheretherketones, polyetherketones, melamine formaldehyde, phenolic formaldehyde, polyamides (e.g., nylon 6, nylon 66, nylon 12), polyamide-imide, polydicyclopentadiene, polyether-imides, polyethersulfones, polyimides, polyphenyleneoxides, polyphthalamide, methylmethacrylate, polyurethanes, polysulfones, poly
  • the plastics used to make the chip include, but are not limited to: polystyrene, polypropylene, polybutadiene, polybutylene, epoxies, TeflonTM, PET, PTFE and chloro-fluoroethylenes, polyvinylidene fluoride, PE-TFE, PE-CTFE, liquid crystal polymers, Mylar®, polyester, LDPE, HDPE, polymethylpentene, polyphenylene sulfide, polyolefins, PVC, and chlorinated PVC.
  • microchannels typically use smaller channels (referred to herein as microchannels or microconduits) than have been used by previous workers in the field.
  • the microchannels (microconduits) according to the claimed invention have widths of 20 ⁇ m, whereas channels an order of magnitude larger have typically been used by others when capillary forces are used to move fluids. Depths of the microchannels are 20 ⁇ m.
  • the resistance to movement can be overcome by a pressure difference, for example, by applying pumping, vacuum, electroosmosis, heating, or additional capillary force. As a result, liquids can move from one region of the device to another as required for the analysis being carried out.
  • the consumable devices of the presently claimed and disclosed inventive concepts are generally small and flat, typically, but not limited to, about 0.5 to 2 square inches (12.5 to 50 mm 2 ) or disks having, but not limited to, a radius of about 15 to 60 mm.
  • the volume of apportioned fluid sample introduced into a particular microfluidic circuit will be small.
  • the sample typically will contain only about 0.1 to 10 ⁇ L for each assay, although the total volume of a specimen may range from 10 to 200 ⁇ L.
  • the consumable of the presently claimed and disclosed inventive concepts comprises a square or rectangular strip or card, or disk.
  • the consumable (chips) used in the presently claimed and disclosed inventive concepts generally are intended to be disposable after a single use.
  • disposable chips will be made of inexpensive materials to the extent possible, while being compatible with the reagents and the samples which are to be analyzed.
  • the test device 10 includes a first well 12 with an on-board reagent and a second well 14 with an on-board calibrator.
  • On-board means that they were placed in the test as part of a manufacturing process rather than at the time of conducting the assay.
  • Each of the first and second wells have a flow path 16 through which the sample mixed with reagent and calibrator, respectively, may flow.
  • the sample is placed in each of the wells via a pipette. Samples 5-50 ⁇ l range with around 20 ⁇ l used in a preferred embodiment consistent with the volume of a traditional finger stick sample.
  • the pipette may be part of an automated or semi-automated analyzer, or may be handled manually by an operator. The metering and mixing necessary for the reaction are handled via the pipette. This reduces the complexity of managing these critical functions on the consumable.
  • the flow paths have a transparent or translucent portion 18. These transparent portions are where the test can be read optically by a detection device.
  • the flow paths are arranged closely to one another and are aligned such that the detection device can capture images from both flow paths simultaneously.
  • the flow paths may end in a vent, well, or aperture connected to a pump 20 to move the fluid through the flow path.
  • an analytical system in accordance with the invention includes a test cartridge and an instrument having a pipetting system, a pump, and a detector.
  • the consumable may be used, for example, for a complete blood count and a white blood cell differential.
  • the consumable has on-board reagents and a calibrator.
  • the reagents may be standard reagents and calibrators known in the art of hematology.
  • the reagent may also include sheath fluid. It is understood that this invention may be used for any analysis that can be read optically by substituting the appropriate reagents and adding additional wells and flow paths, if necessary.
  • the consumable may be foil sealed across top.
  • a sample is loaded into sample well A.
  • An instrument 30 dispenses metered sample in wells B and C utilizing an automated pipette 34.
  • the pipette may be on a track to access multiple wells.
  • Wells B and D contain Staining reagents for RBC and WBC's.
  • Example stains include Eosin or Wright's stains.
  • Well C is contains a cell lysis reagent. Several commercial lysis reagents are commonly available such as EasySep or Roche. A fixed volume of sample is transferred from well C to well D for staining utilizing the pipette.
  • Well E contains calibrator.
  • the calibrator may consist of precise volume of particles (fluorescent or colored) that can be used to normalize dimensional errors in manufacturing.
  • the particles are highly precise in concentration and size distribution and are typically polystyrene from commercial vendors such as Polysciences or Spherotech.
  • flow commences through a 3-channel array using an external pump on the instrument.
  • the pump 18 may be connected to a pressure sensor and a feedback control.
  • the pump for example, may be a syringe pump, a peristaltic pump, a piezoelectric pump, or the like, which provides a required flow rate.
  • the connecting element for connecting the pump to the consumable may be a tube or hose.
  • the Field of View (FOV) for the imager's 36 high- objective lens must accommodate simultaneous imaging. Typical magnification ranges from 10-40x with the working length being dependent on the type of objective used. Images are captured on conventional imagers such as a CCD or CMOS imager that can capture the desired FOV and has the resolution to adequately discriminate the particles. These images are conventionally available from commercial vendors. Images are captured through precise apertures that define the FOV with high accuracy. This allows normalizing the field of view with the calibrator reducing the sensitivity to the depth.
  • the primary impact of a variable depth in the microchannel is to change the concentration of the particles relative to the buffer - i.e., the viewed volume contains a different volume of the original sample from the nominal depending on the change in depth (note that the other two dimensions are controlled by the precision aperture). Since the calibrator exhibits the same variability, however with a well-known concentration, it can be used to normalize the impact of depth. Specialized analytical software is then used to analyze the image utilizing the known volume of the sample calculated utilizing the dimensions of the pinhole apertures and the calibrant to quantify the analyte in the bodily fluid sample. The instrument can then print out the result on a screen, onto paper, or export the data into an informatics system or data collection unit.
  • the first step is providing a consumable in accordance with the invention described above.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Fluid Mechanics (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Claims (3)

  1. Testvorrichtung, umfassend:
    a. einen ersten Well mit einem darin angeordneten Reagens;
    b. einen zweiten Well mit einer darin angeordneten Kalibriersubstanz; und
    c. einen Fließweg von sowohl dem ersten als auch dem zweiten Well mit einem transparenten Abschnitt,
    wobei der Fließweg als Mikrokanäle mit Breiten von 20 µm und Tiefen von 20 µm ausgestaltet ist.
  2. Testvorrichtung nach Anspruch 1, wobei die transparenten Abschnitte der Fließwege nahe beieinander angeordnet und derart ausgerichtet sind, dass eine Detektionsvorrichtung Bilder aus beiden Fließwegen gleichzeitig aufnehmen kann.
  3. Analysesystem, umfassend:
    d. die Testvorrichtung nach Anspruch 1; und
    e. ein Instrument, das die Testvorrichtung aufnehmen kann und umfasst:
    i. eine Pipette;
    ii. einen optischen Leser mit einer oder mehreren Öffnungen, die derart strukturiert und angeordnet sind, dass gleichzeitig Objekte in den transparenten Abschnitten von wenigstens zwei Fließwegen an der Testvorrichtung abgebildet werden können; und
    iii. eine Pumpe, die mit der Testvorrichtung verbindbar ist.
EP15816063.0A 2014-06-30 2015-06-29 Mikrofluidische testkartusche ohne aktive fluidregelung Active EP3160647B1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201462018890P 2014-06-30 2014-06-30
PCT/US2015/038361 WO2016003927A1 (en) 2014-06-30 2015-06-29 Microfluidic test cartridge with no active fluid control

Publications (3)

Publication Number Publication Date
EP3160647A1 EP3160647A1 (de) 2017-05-03
EP3160647A4 EP3160647A4 (de) 2017-07-26
EP3160647B1 true EP3160647B1 (de) 2021-07-28

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EP15816063.0A Active EP3160647B1 (de) 2014-06-30 2015-06-29 Mikrofluidische testkartusche ohne aktive fluidregelung

Country Status (3)

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US (1) US20170197212A1 (de)
EP (1) EP3160647B1 (de)
WO (1) WO2016003927A1 (de)

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US11717823B2 (en) 2020-01-06 2023-08-08 Bisu, Inc. Microfluidic system, device and method
AT525192A1 (de) * 2021-06-15 2023-01-15 Genspeed Biotech Gmbh Mikrofluidischer chip

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WO1999060397A1 (en) * 1998-05-18 1999-11-25 University Of Washington Liquid analysis cartridge
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Also Published As

Publication number Publication date
EP3160647A4 (de) 2017-07-26
US20170197212A1 (en) 2017-07-13
WO2016003927A1 (en) 2016-01-07
EP3160647A1 (de) 2017-05-03

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